Anti-CD44v6抗体[VFF-18] (ab78960)

CD44v6 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

ab78960 (Ascites) staining CD44v6 in human breast cancer tissue by Immunohistochemistry (Frozen sections).Tissue was fixed in methanol and then blocked using 10% serum for 5 minutes at 25°C. Samples were then incubated with ab78960 at a 1/500 dilution for 1 hour at 25°C. The secondary used was an undiluted HRP conjugated goat polyclonal.

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Overlay histogram showing THP1 cells stained with ab78960 (Ascites) (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78960, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Immunocytochemical analysis of methanol fixed cultured human breast epithelial organoids labeling CD44v6 with ab78960 (Ascites) at 1/500 dilution. UltraVision ONE HRP Polymer was used as the secondary antibody. 10% serum was used as the blocking agent. 

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IHC image of ab78960 (Ascites) staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78960, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.